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A novel three-dimensional culture system allows prolonged culture of functional human granulosa cells and mimics the ovarian environment


Kossowska-Tomaszczuk, K; Pelczar, P; Güven, S; Kowalski, J; Volpi, E; De Geyter, C; Scherberich, A (2010). A novel three-dimensional culture system allows prolonged culture of functional human granulosa cells and mimics the ovarian environment. Tissue Engineering. Part A, 16(6):2063-2073.

Abstract

The development of techniques allowing the growth of primordial follicles to mature follicles in vitro has much potential for both reproductive medicine and developmental research. However, human primordial and preantral follicles fail to grow after isolation from the surrounding ovarian stroma. Granulosa cells (GCs), which normally undergo apoptosis after ovulation, contain a subpopulation of ovarian follicular cells remaining viable in vitro over prolonged time periods when cultured in the presence of leukaemia inhibiting factor (LIF). However, when cultured as monolayers, they progressively lose all their characteristics, such as follicle-stimulating hormone receptor (FSHR) and cytochrome P450-aromatase. Here, we describe a three-dimensional (3D) culture system containing type I collagen, which, together with LIF, allowed the survival and growth of a subpopulation of GCs isolated from mature ovarian follicles and supported them to proliferate into spherical structures exhibiting steroidogenic capacity, as demonstrated by P450-aromatase and 3-hydroxysteroid dehydrogenase. After transplantation into the ovaries of immuno-deficient mice, these cells became localized preferentially within antral follicles and the prolonged expression of FSHR was confirmed as well. With this optimization of the culture conditions an environment was created, which acts as a niche closely mimicking the development of early ovarian follicles in vitro.

The development of techniques allowing the growth of primordial follicles to mature follicles in vitro has much potential for both reproductive medicine and developmental research. However, human primordial and preantral follicles fail to grow after isolation from the surrounding ovarian stroma. Granulosa cells (GCs), which normally undergo apoptosis after ovulation, contain a subpopulation of ovarian follicular cells remaining viable in vitro over prolonged time periods when cultured in the presence of leukaemia inhibiting factor (LIF). However, when cultured as monolayers, they progressively lose all their characteristics, such as follicle-stimulating hormone receptor (FSHR) and cytochrome P450-aromatase. Here, we describe a three-dimensional (3D) culture system containing type I collagen, which, together with LIF, allowed the survival and growth of a subpopulation of GCs isolated from mature ovarian follicles and supported them to proliferate into spherical structures exhibiting steroidogenic capacity, as demonstrated by P450-aromatase and 3-hydroxysteroid dehydrogenase. After transplantation into the ovaries of immuno-deficient mice, these cells became localized preferentially within antral follicles and the prolonged expression of FSHR was confirmed as well. With this optimization of the culture conditions an environment was created, which acts as a niche closely mimicking the development of early ovarian follicles in vitro.

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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:05 Vetsuisse Faculty > Institute of Laboratory Animal Science
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:January 2010
Deposited On:03 Mar 2010 14:00
Last Modified:05 Apr 2016 14:02
Publisher:Mary Ann Liebert
ISSN:1937-3341
Publisher DOI:10.1089/ten.TEA.2009.0684
PubMed ID:20109057
Permanent URL: http://doi.org/10.5167/uzh-32724

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